{
  "abstract": "Mass spectrometry (MS)-based single-cell proteomics (SCP) enables proteome-wide analysis at single-cell resolution, offering insights into cellular heterogeneity, biological processes, and disease mechanisms. However, conventional nanoLC-MS workflows are constrained by long gradients and low throughput, limiting their application to small cohorts. Here, we present a multicolumn nanoLC-MS platform that achieves 5-minute separation windows with 100% duty cycles at ~100 nL/min, enabling the analysis of up to 288 single cells per day. The system provides stable peptide separation, negligible carryover, and robust retention-time reproducibility across nearly 1200 consecutive injections. We successfully analyze more than 4000 samples at nearly 288 samples per day (SPD) throughput. The platform identifies ~5000 proteins per 250 pg digest injection and an average of ~4000 proteins per single HeLa cell with maxima exceeding 5100, which is comparable to state-of-the-art longer-gradient workflows. Quantitative benchmarking with mixed-species standards confirm the quantitative accuracy is largely maintained during extended acquisition. Applying the workflow to 783 LPS-treated RAW264.7 macrophages reveals substantial cellular heterogeneity under a uniform inflammatory stimulus. Together, these results establish a robust and scalable platform for high-throughput SCP, enabling thousands of single-cell analyses within a single study while maintaining deep proteome coverage and biological interpretability.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA",
    "Brigham Young University",
    "MicrOmics Technologies",
    "MicrOmics Technologies, LLC, Spanish Fork, UT, USA",
    "Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA. ryan.kelly@byu.edu"
  ],
  "article_type": "research support, non u.s. gov't",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Chao Wang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Hsien‐Jung L. Lin",
      "orcid": "0000-0003-4704-3692"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Siqi Huang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Garrett Haynie",
      "orcid": "0009-0004-4547-5330"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Kenneth Triggs",
      "orcid": "0009-0001-5091-7058"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Ella E. Norton"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "R. J. S. Miercort"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yen-Jou Chang"
    },
    {
      "affs": [
        0,
        2
      ],
      "name": "David V. Hansen"
    },
    {
      "affs": [
        3,
        2
      ],
      "name": "Thy Truong"
    },
    {
      "affs": [
        3
      ],
      "name": "Xiaofeng Xie",
      "orcid": "0000-0002-6731-7796"
    },
    {
      "affs": [
        4
      ],
      "name": "Ryan Kelly",
      "orcid": "0000-0002-3339-4443"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/egkx120ay1r0q.bib",
  "cited_by": 8,
  "doi": "10.1038/s41467-026-78059-x",
  "fulltext_url": "https://europepmc.org/articles/PMC13691111",
  "id": "egkx120ay1r0q",
  "issns": [
    "2041-1723"
  ],
  "landing_url": "https://europepmc.org/article/MED/42918804",
  "language": "eng",
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1607|Advanced Proteomics Techniques and Applications"
    ]
  ],
  "openalex": "W4416322320",
  "pdf_url": "https://europepmc.org/articles/PMC13691111?pdf=render",
  "pmcid": "PMC13691111",
  "pmid": "42918804",
  "published": "2026-09-23",
  "publisher": "Nature Portfolio",
  "ris": "https://research.draconitic.com/paper/egkx120ay1r0q.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:20:36Z",
      "native_id": "MED:42918804",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42918804"
    },
    {
      "first_seen": "2026-10-30T13:26:44Z",
      "native_id": "W4416322320",
      "source": "openalex",
      "url": "https://doi.org/10.1038/s41467-026-78059-x"
    }
  ],
  "subjects": [
    {
      "field": "chemistry",
      "origin": "openalex:1607|Advanced Proteomics Techniques and Applications",
      "subfield": "physical"
    }
  ],
  "title": "High-throughput label-free single-cell proteomics enabled by multicolumn NanoLC.",
  "title_text": "High-throughput label-free single-cell proteomics enabled by multicolumn NanoLC.",
  "type": "peer-reviewed",
  "updated_at": "2026-11-09T13:21:03Z",
  "url": "https://research.draconitic.com/paper/egkx120ay1r0q",
  "venue": "Nature communications",
  "versions": []
}